基于UHPLC-Q-Orbitrap HRMS比较古今不同加工方法对百合煎液化学成分的影响

Comparison of Effect of Ancient and Modern Processing Methods on Chemical Components of Lilii Bulbus Decoction Based on UHPLC-Q-Orbitrap HRMS

目的:比较古今不同加工方法对百合煎液化学成分的影响,为该药材的产地加工、饮片炮制及临床应用提供依据。方法:采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法(UHPLC-Q-Orbitrap HRMS),利用准分子离子、特征碎片离子等信息,并参考相关文献和数据库信息进行化合物结构鉴定;利用主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)筛选主要差异成分,通过高效液相色谱法对差异成分进行定量研究,比较百合不同加工品煎液中化学成分种类和含量的区别。结果:从百合不同加工品煎液及鲜百合水浸液、烫煮液中共鉴别出24个成分,其中17个酚类、5个皂苷类、2个生物碱类;与鲜百合煎液比较,烫煮干百合煎液中王百合苷A、对香豆酸、秋水仙碱等成分含量减少,蒸制干百合煎液中王百合苷C含量降低,水浸鲜百合煎液中王百合苷A、王百合苷C成分含量减少;与烫煮干百合煎液比较,鲜百合煎液与水浸鲜百合煎液中成分的整体含量要高,蒸制干百合煎液除王百合苷C的含量略低外,其他成分含量要高于烫煮干百合煎液。结论:不同加工过程会对百合煎液化学成分的种类及含量产生一定影响,烫煮法加工鲜百合有利于百合的保存,但会造成百合有效成分流失;相较于烫煮法,蒸制法高温处理可防止百合褐变,还能降低其有效成分流失;张仲景提出水浸一宿后去沫的加工方法可能更有利于治疗百合病等温热病,可为百合不同加工品的临床合理应用及作用机制研究提供参考。

Objective:To compare the effects of different processing methods in ancient and modern times on the chemical components of Lilii Bulbus decoction,and to provide experimental support for the origin processing,decoction piece processing and clinical application of this herb.Method:Ultra high performance liquid chromatography tandem quadrupole electrostatic field orbitrap high resolution mass spectrometry(UHPLCQ-Orbitrap HRMS)was used for structural identification of the compounds using excimer ions,secondary MS and characteristic fragment ions,and referring to relevant literature and database information.Principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were used to screen the main differential components,the differential components were quantitatively studied by high performance liquid chromatography(HPLC),in order to compare the types and contents of chemical components in the decoction of different processing products of Lilii Bulbus.Result:A total of 24 chemical components were identified from the decoction of different processed products of Lilii Bulbus,water extract and scalding liquid of fresh Lilii Bulbus,including 17 phenols,5 saponins and 2 alkaloids.Compared with the fresh Lilii Bulbus decoction,the contents of regaloside A,p-coumaric acid,colchicine and other components in the decoction of dry Lilii Bulbus processed by scalding method decreased,the content of regaloside C in the decoction of dry Lilii Bulbus processed by steaming method decreased,and the contents of regaloside A and regaloside C in the decoction of fresh Lilii Bulbus processed by water immersion also decreased.Compared with the decoction of dry Lilii Bulbus processed by scalding method,the overall content of components in the fresh Lilii Bulbus decoction and the decoction of fresh Lilii Bulbus processed by water immersion was higher,the contents of components in the decoction of dry Lilii Bulbus processed by steaming method was higher,except for the slightly lower content of regaloside C.Conclusion:Different processing processes have a certain effect on the types and contents of chemical components in Lilii Bulbus decoction.Scalding process is beneficial to the preservation of Lilii Bulbus,but can cause the loss of effective components.Compared with scalding method,steaming method can prevent browning of Lilii Bulbus and reduce the loss of its active ingredients.The processing method of removing foam after overnight immersion proposed by ZHANG Zhongjing may be more conducive to the treatment of Baihe disease,which can provide reference for the clinical rational application and mechanism research of different processed products of Lilii Bulbus.